CN107607631A - A kind of method of lycoramine content in quick detection lycoris plants - Google Patents
A kind of method of lycoramine content in quick detection lycoris plants Download PDFInfo
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Abstract
The present invention relates to a kind of method of lycoramine content in quick detection lycoris plants, including：(1) analyte sample fluid is prepared；(2) detect；(3) cubage.The present invention can carry out rapid extraction and qualitative and quantitative analysis to the alkaloids substance lycoramine in lycoris plants, analysis time is greatly shortened, it can complete to detect program within 6 minutes, lycoramine retention time is within 3.6 minutes, quickly obtain testing result, analyze speed and accuracy are substantially increased, efficiently and accurately analysis method is provided for the accumulation law and anabolism regulation and control of lycoramine in research lycoris plants.
Description
Technical field
The invention belongs to alkaloids substance detection field, Li Kela in more particularly to a kind of quick detection lycoris plants
The method of quick content.
Background technology
Lycoris (Lycoris spp.) is Amaryllidaceae (Amaryllidaceae) napiform root herbaceous plant, and many kinds are China
It is distinctive to view and admire and medical flower, rich in the distinctive alkaloid component of a variety of lycoris plants.So far, from lycoris plants
In isolate the alkaloid compound more than 500 kinds of configurations, fitochemical studies show that this kind of compound has a variety of
Pharmacological effects, medical value are high.Wherein, lycoramine treatment senile dementia, sequela of poliomyelitis paralysis,
Play an important role in terms of ischiatitis (Cortes et al., 2015).So far, wild short-tube lycoris resource is still medicinal power
Can the most important source of stretching-sensitive, therefore, the content of lycoramine in short-tube lycoris is carried out accurate qualitative and quantitative analysis be breeding and
One of important content of alkaloids substance anabolism research.
At present, mainly there is TLC separation technology (thin to the method that lycoramine content is analyzed in short-tube lycoris
Layer chromatography, TLC), high performance liquid chromatography detection technology (high-performance liquid
Chromatography), gas chromatography-mass spectrography analytical technology (Gas Chromatography-Mass
) and high performance liquid chromatography-electro-spray ionization-mass spectrometry analytical technology (HPLC-electrospray Spectrometry
ionization-mass spectrometry,HPLC-ESI-MS)。
Li Mingkai (research of foundation and its biosynthesis of maryllidaceous alkaloid analysis method, Agricultural University Of Anhui, 2012：
TLC separation technology 1-69) is utilized, realizes the initial gross separation of the alkaloids substances such as lycoramine, and it was found that two-way
Thin-layer chromatography is more suitable for the separation of a variety of different alkaloids substances than one dimension chromatography, this method mainly by with known biology
The mobility of alkali standard specimen (galanthamine, lycoramine, lycorine) is contrasted, realize in sample alkaloid species it is preliminary
It is qualitative, but cannot be used for accurate structure and quantitative analysis.
Yuan Juhong etc. (learn by Yuan Juhong etc., Lycoris difference inter-species alkaloid Difference, Agricultural University Of Jiangxi
Report, 32 (3)：0560-0565,2010) high performance liquid chromatography detection technology is utilized, pass through sample and known alkaloid standard items
The contrast of retention time and peak area under similarity condition, galanthamine, lycoramine in Lycoris difference plant are carried out
With the quantitative study of lycorine, its single-trial extraction short-tube lycoris bulb material needs fresh sample 60g (water content 90% or so), converts into short-tube lycoris
Bulb dry sample 6g is, it is necessary to consume ethyl acetate 75ml, sodium hydroxide 10ml, methanol 8ml, and pole is consumed to plant sample and reagent
Greatly, meanwhile, it carries out content detection according to the retention time of material, test substance can not be carried out only with HPLC method
It is accurate qualitative, therefore material quantitative result is unreliable, it is impossible to carry out structure, the analysis of qualitative and accurate quantification.
(Chun et al., the Isolation and identification of alkaloids and such as Chun
anthocyanins from flower and bulb of Lycoris radiata using HPLC and LC-ESI-
MS, Journal of Agricultural Chemistry and Environment, 2 (1)：22-26,2013) use
HPLC-ESI-MS method, quantitative analysis being carried out to the lycoramine in short-tube lycoris, the elution time of lycoramine is 46.9min,
HPLC analysis times are relatively long, it is necessary to 60 minutes, and it carries out content detection, the method according to retention time and parent ion information
The defects of compared with Yuan Juhong etc. method, accuracy has been lifted, but obvious be, the thing of identical elution time and parent ion size
Matter composition may have many kinds, therefore still defective in qualitative, quantitative accuracy.
Also, extraction time length in the prior art, also be present, it is cumbersome the problem of, it is main in such as Yuan Juhong method
Want extraction step for drying, crush, sieving, wetting, ethyl acetate extraction, backflow in water-bath, extract solution filter out, ethyl acetate water
Flowed back in bath, filter out flowed back in extract solution, ethyl acetate water-bath, filter out extract solution, merge backflow extract solution, rotation be evaporated,
The steps such as cooling, methanol washing dissolving, filtering, make troubles for the fast quantitative analysis of lycoramine.
The content of the invention
The technical problems to be solved by the invention are to provide lycoramine content in a kind of quick detection lycoris plants
Method, this method can carry out rapid extraction and qualitative, quantitative point to the alkaloids substance lycoramine in lycoris plants
Analysis, analysis time is greatly shortened, can be completed within 6 minutes detection program, lycoramine retention time 3.6 minutes with
It is interior, testing result is quickly obtained, substantially increases analyze speed and accuracy, for the product of lycoramine in research lycoris plants
Tired rule and anabolism regulation and control provide efficiently and accurately analysis method.
The invention provides a kind of method of lycoramine content in quick detection lycoris plants, including：
(1) lycoris plants sample to be measured is weighed, is pulverized after freeze-drying, extraction is used as using alcohol organic solvent
Agent, ultrasonic extraction, mixing, centrifugation, merge supernatant, concentration, then redissolve, be diluted to 1~100ng/ml of concentration, it is molten to obtain testing sample
Liquid；
(2) testing sample solution is detected using liquid chromatography-tandem second order mses, obtains lycoramine in sample
Peak area data；
Wherein, chromatographic condition is as follows：
Liquid-phase chromatographic column：Silica gel type reverse-phase chromatographic column；
Column temperature：40~50 DEG C；
Sample size：5~10 μ L；
Flow velocity：0.2~0.4ml/min；
Mobile phase：Mobile phase A：Formic acid-the aqueous solution, Mobile phase B：Acetonitrile or methanol；
Elution process：The volume ratio of gradient elution, elution time≤6.0min, mobile phase A and Mobile phase B is 40~95：5
~60；
Mass Spectrometry Conditions and scan mode：
Ion gun：Heat electric spray ion source or electric spray ion source；
Ion gun spray voltage：2500~4500V；
Scan mode：With first mass spectrometric full scan-data dependence daughter ion scan pattern or more reactions under cation scanning
Detection pattern is analyzed；
Monitor ion pair：290 ± 1Da of parent ion m/z and at least one lycoramine daughter ion；
Lycoramine retention time：≤3.6min；
(3) lycoramine in sample solution is obtained according to the standard curve of the peak area data of step (2) and lycoramine
Concentration, in conjunction with solvent volume and sample dry weight is redissolved in step (1), calculate the content of lycoramine in sample.
Alcohol organic solvent in the step (1) is methanol or ethanol, and the ratio of sample and extractant is 0.2-1g:2-
5ml。
The ultrasonic extraction time in the step (1) is 15-40min.
The solvent of redissolution in the step (1) is mobile phase primary condition, i.e. 95vol% water -5vol% acetonitriles, is contained
0.1v/v% formic acid.
The formic acid content in formic acid-aqueous solution in the step (2) is 0.1~1v/v%.
The mass-to-charge ratio m/z scopes of daughter ion in the step (2) are 58~290Da.
Preferably, the mass-to-charge ratio m/z of the daughter ion is 233 ± 1Da, 215 ± 1Da or 189 ± 1Da.
Preferably, the chromatographic condition in the step (2) and Mass Spectrometry Conditions are：40 DEG C of column temperature, the μ L of sample size 5~10, stream
0.2~0.4ml/min of speed, ion gun are heating electric spray ion source, and scan mode is cation Full MS/dd-MS2, ion
Source spray voltage 2500V, elution time 6.0min, the retention time of lycoramine is 3.54~3.55min.
The preparation method of the standard curve of lycoramine in the step (3) is：Lycoramine standard items are taken, are configured to
Concentration 1ng/ml~100ng/ml series standard solution, it is molten to lycoramine standard using liquid chromatography-tandem second order mses
Liquid is measured, and is drawn standard curve using the peak area data and lycoramine concentration of standard solution measured, is obtained linear side
Journey.
Chromatographic condition and the Mass Spectrometry Conditions for determining lycoramine standard liquid are identical with step (2).
The present invention using high performance liquid chromatography-heating electric spray ion source-second order mses (HPLC-HESI-MS/MS) technology as
Basis, gradient elution is carried out, by analysis time by prior art high performance liquid chromatography-electric spray ion source-first mass spectrometric
(HPLC-ESI-MS) 60 minutes of method were foreshortened within 6 minutes, and second order mses of having connected, analyze lycoramine from
The information of son, confirms through methodology, achieves extraction and analytical effect well, substantially increases analyze speed and accuracy,
A kind of efficiently and accurately analysis method is provided to study the accumulation law of Lycoris lycoramine and anabolism regulation and control.
For the present invention when handling testing sample, the extract solution of selection is safer, both can guarantee that analytical effect while or can
To improve the security of operation.Meanwhile when being extracted, the plant sample and amount of reagent of consumption are few, and testing sample fresh weight can be low
To 0.2g, extractant volume is 2-5ml.On the premise of extraction effect is ensured, vegetable material and extracts reagent have greatly been saved.
Also, the present invention takes quickly simplified extracting method, 15-40min is extracted by more than 2 times ultrasonic assistants, centrifuges, steams
Hair, redissolve, filtering, be computed, the rate of recovery is up to 95% or so, and extraction process is quick and easy, and the rate of recovery is high.
The present invention is in lycoramine content in detecting testing sample, in the range of certain retention time, without other
The interference of characteristic ion pair, therefore, using the gradient elution mode of the present invention, shorten detection program time, detect program time
≤ 6min, elution time≤3.6min of lycoramine, preferably 3.54-3.55min, improves detection speed.
The present invention is in detection, liquid chromatography tandem second order mses, by the retention time of material, parent ion, son from
The three-level information integration of son, the information monitoring of multigroup parent ion/daughter ion pair particularly in second order mses, it can ensure that qualitative and fixed
The accuracy of amount.
In preferred embodiments of the present invention, using heating electric spray ion source (HESI), determinand cation scanning under with
Full MS/dd-MS2 patterns are analyzed, and are scanned in the positive-ion mode, parent ion [M+H]+M/z 290 response
Highest, the higher two daughter ions m/z215 and m/z189 of reselection relative abundance are monitoring ion, can greatly improve sensitivity.
Beneficial effect
The present invention can carry out rapid extraction and qualitative, quantitative to the alkaloids substance lycoramine in lycoris plants
Analysis, analysis time is greatly shortened, can be completed within 6 minutes detection program, lycoramine retention time 3.6 minutes with
It is interior, testing result is quickly obtained, the lycoramine parent ion and daughter ion information monitoring provided by second order mses, it is fixed to improve
Property and quantitative analysis accuracy, the rate of recovery of lycoramine is 94.8~96.1%, relative standard deviation (RSD) is 1.0~
6.5%, separating degree realizes the quick elution separation of lycoramine to be kept completely separate, can detect multiple parent ions of lycoramine/
Daughter ion is to information, it is ensured that qualitative and quantitative accuracy, accumulation law and conjunction for lycoramine in research lycoris plants
Efficiently and accurately analysis method is provided into metabolic regulation.
Brief description of the drawings
Fig. 1 is the standard curve of the lycoramine in the embodiment of the present invention 1；
Fig. 2 is the chromatogram of the lycoramine in the embodiment of the present invention 1；
Fig. 3 is the MS2 mass spectrograms of the lycoramine in the embodiment of the present invention 1.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.In addition, it is to be understood that after the content of the invention lectured has been read, people in the art
Member can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
Embodiment 1
1) standard curve is drawn
Lycoramine standard items 0.010g (Lycm, Lycoramine, C are weighed with assay balance precision17H23NO3, molecular weight
289.375), with methanol dissolving, simultaneously constant volume to 100ml, obtains its standard reserving solution, Stock concentrations are 100 μ g/mL；
Interstitial fluid in standard：Accurately pipette 1mL standard reserving solutions to be placed in 100mL volumetric flasks, 1.0 μ are made into methanol constant volume
Interstitial fluid in g/mL hybrid standard；
Standard working solution：Pipette interstitial fluid in appropriate hybrid standard as needed, respectively with methanol dilution into concentration 1ng/mL,
5ng/mL, 10ng/mL, 50ng/mL and 100ng/mL standard working solution.
Lycoramine standard liquid is measured using the chromatograph for second order mses instrument of having connected, utilizes the peak face measured
Volume data and lycoramine concentration of standard solution draw standard curve, linear equation are obtained, referring to Fig. 1.
As a result show that lycoramine its mass concentration and peak area in the range of 1ng/ml-100ng/ml are in good line
Sexual intercourse, linear relationship equation are Y=632925X, coefficient R2=0.9981, detection limit and quantification range are as shown in table 1：
Table 1
With it is actually detected when S/N=3 determine that the detection of method is limited to 0.2ng/g, the detection of instrument is limited to 0.1ng/ml.
2) testing sample is handled
Using the bulb for changing brocade flower as testing sample 0.2g (water content 90% or so), after freeze-drying, pulverize, obtain stone
Garlic bulb dry sample 0.02g, take short-tube lycoris bulb dry sample to be placed in EP pipes after mixing and add the ethanol of 2mL 70%, ultrasonic extraction 28min,
12000rpm centrifuges 10min, takes supernatant, and filter residue adds the ethanol of 1.5mL 70% ultrasonic extraction 28min, vortex 1min again,
12000rpm centrifuges 10min, takes supernatant, merges supernatant twice, and nitrogen, which blows, is concentrated near dry, 1mL mobile phase primary condition
(95vol% water -5vol% acetonitriles, formic acid containing 0.1v/v%) redissolves, and D times of dilution (being 100 times in the present embodiment) to concentration 1~
100ng/ml, cross 0.22 μm of filter membrane.
3) chromatogram detects
Utilize ultra performance liquid chromatography-thermoelectricity esi ion source-second order mses Combined techniques (UHPLC-HESI-MS/
MS), testing sample is detected, obtains the peak area data of lycoramine in sample；
Wherein, chromatographic condition：
Liquid-phase chromatographic column：Waters BEH C18(2.1x 100mm,1.7μm)；
Column temperature：40℃；
Sample size：5μL；
Flow velocity：0.2ml/min；
Mobile phase：Mobile phase A：Formic acid volume fraction be 0.1% formic acid-aqueous solution, Mobile phase B：Acetonitrile；
Elution requirement：Gradient elution, elution program are as follows：
Table 2
Chromatogram is referring to Fig. 2, from Figure 2 it can be seen that under these conditions, lycoramine elution time is 2.58min, and in journey
In sequence analysis time, disturbed without other characteristic peaks, can be achieved to be kept completely separate.
Mass Spectrometry Conditions：
Mass spectrum is using Thermo ScientificTMQ ExactiveTMMass Spectrometer Method system, equipped with electron spray
(ESI) ion gun and Xcalibur work stations.
Ion gun：Heat electric spray ion source (HESI)；
Scan mode：Analyzed under cation scanning with Full MS/dd-MS2 patterns；
First mass spectrometric resolution ratio is 70000, two level 35000；
Ion gun spray voltage 2500V；
350 DEG C of capillary temperature；
Gas：Nitrogen；Sheath throughput：10；Secondary air amount：2；
Purge gas velocity：0；
S- lens are horizontal：55；
40 DEG C of heter temperature；
Monitor ion pair：290.1/215.1 and 290.1/189.1, referring to table 3；
Retention time is 3.54min；
The target compound ion information of table 3
Mass spectrogram is referring to Fig. 3, as seen from Figure 3, parent ion [M+H]+M/z 290 response highest, relative abundance are higher
Two daughter ions m/z215 and m/z189 for monitoring daughter ion.
4) cubage
By in the linear equation obtained by the mass spectra peak area data substitution step 1) of testing sample, it is molten to obtain testing sample
The concentration C (unit ng/ml) of lycoramine in liquid, it is dilute further according to the mobile phase primary condition volume 1ml redissolved in step 2)
Multiple D, and sample quality 0.02g are released, brings cubage formula I into, calculating the content m of lycoramine in sample, (unit is
Ng/g dry weights).
Cubage formula I is：M=(C × V × D)/M.
Wherein, lycoramine content in m=samples, unit are ng/g dry weights；Lycoramine concentration in C=solution to be measured,
Unit ng/ml；The solvent volume that V=redissolves, unit ml；D=extension rates；M=sample qualities, dry weight, unit g.
Such as certain part determined in this example changes brocade flower bulb sample, C=20.353ng/ml, V=1ml, D=100, M=
0.0265g, calculated according to formula and finally drawn：M=76802ng/g dry weights.
The weight pulverized after testing sample freeze-drying is dry weight.
Further, recovery testu is carried out using the bulb sample for changing brocade flower, is added under 6 μ g/ml pitch-based spheres
Determination of recovery rates is marked, according to step 2-3) method carry out, each level is repeated 3 times, and is as a result measured to and changes brocade flower bulb sample
Between 76802~108280ng/g (i.e. 0.076~0.108mg/g) dry weight, the rate of recovery is the content of middle lycoramine
96.1%, RSD 6.5%, show established extraction and analytical method accurately and reliably.
Embodiment 2
1) standard curve is drawn
Lycoramine standard items 0.010g (Lycm, Lycoramine, C are weighed with assay balance precision17H23NO3, molecular weight
289.375), with methanol dissolving, simultaneously constant volume to 100ml, obtains its standard reserving solution, Stock concentrations are 100 μ g/mL；Standard
Middle interstitial fluid：Accurately pipette 1mL standard reserving solutions to be placed in 100mL volumetric flasks, 1.0 μ g/mL mixing mark is made into methanol constant volume
Interstitial fluid in standard；Standard working solution：Interstitial fluid in appropriate hybrid standard is pipetted as needed, respectively with methanol dilution into concentration 1ng/
ML, 2ng/mL, 5ng/mL, 25ng/mL, 50ng/mL standard working solution.
Lycoramine standard liquid is measured using the chromatograph for second order mses instrument of having connected, utilizes the peak face measured
Volume data and lycoramine concentration of standard solution draw standard curve.
As a result show that lycoramine its mass concentration and peak area in the range of 1ng/ml-100ng/ml are in good line
Sexual intercourse, linear relationship equation are Y=1.0e6X-126075, coefficient R2=0.9994, detection limit and quantification range such as table 4
It is shown：
Table 4
With it is actually detected when S/N=3 determine that the detection of method is limited to 0.2ng/g, the detection of instrument is limited to 0.1ng/ml.
2) testing sample is handled
Using the bulb of lycoris plants Lycoris as testing sample, after testing sample freeze-drying, pulverize, mix
After take 0.02g to add the ethanol of 2mL 70% in EP pipes, ultrasonic extraction 28min, 12000rpm centrifugation 10min, take supernatant, filter residue
The ethanol of 1.5mL 70% ultrasonic extraction 28min again is added, vortex 1min, 12000rpm centrifugation 10min, supernatant is taken, merges two
Secondary supernatant, nitrogen, which blows, is concentrated near dry, 1mL mobile phases primary condition (95vol% water -5vol% acetonitriles, formic acid containing 0.1v/v%)
Redissolve, D times of dilution (being 100 times in the present embodiment) crosses 0.22 μm of filter membrane, obtain testing sample solution to 1~100ng/ml of concentration.
3) chromatogram detects
Testing sample is detected using the liquid chromatogram for second order mses instrument of having connected, obtains lycoramine in sample
Peak area data；
Wherein, chromatographic condition：
Liquid-phase chromatographic column：Waters BEH C18(2.1x 100mm,1.7μm)；
Column temperature：50℃；
Sample size：10μL；
Flow velocity is 0.4ml/min；
Mobile phase：Mobile phase A：Formic acid volume fraction be 0.1% formic acid-aqueous solution, Mobile phase B：Methanol；
Elution requirement：Gradient elution, elution program are as follows
Table 5
| Time(min) | Flow rate(mL/min) | %A | %B |
| 0 | 0.4 | 95 | 5 |
| 3.5 | 0.4 | 40 | 60 |
| 4.5 | 0.4 | 40 | 60 |
| 4.51 | 0.4 | 95 | 5 |
| 6 | 0.4 | 95 | 5 |
Under these conditions, lycoramine elution time is 2.50min, and within program analysis time, without other features
Peak disturbs, and can be achieved to be kept completely separate.
Mass Spectrometry Conditions：
Mass spectrometer system using American AB Sciex companies the Mass Spectrometer Method systems of Qtrap 6500, equipped with electron spray
(ESI) ion gun and Analyst 1.6.2 work stations.
Ion gun：Electric spray ion source (ESI)；
Scan mode：Analyzed under cation scanning with more reaction detections (MRM) pattern；
Ion gun spray voltage 4500V；
Temperature：550℃；
Gas curtain gas 30；
Ion gun Gas1：55；Gas2：55；
Monitor ion pair：And 290.3/189.1 290.3/215.1；
Retention time is 3.55min；
4) cubage
By in the linear equation obtained by the mass spectra peak area data substitution step 1) of testing sample, obtain in solution to be measured
The concentration C (unit ng/ml) of lycoramine, further according to methanol volume 1ml, the extension rate D redissolved in step 2), and sample
Quality 0.02g, bring cubage formula I into, calculate the content m of lycoramine in sample (unit is ng/g dry weights).
Cubage formula I is：M=(C × V × D)/M.
Wherein, lycoramine content in m=samples, unit are ng/g dry weights；Lycoramine concentration in C=solution to be measured,
Unit ng/ml；The solvent volume that V=redissolves, unit ml；D=extension rates；M=sample qualities, dry weight, unit g.
Such as determine certain part of Lycoris bulb sample in this example, C=5.684ng/ml, V=1ml, D=100, M
=0.0204g, calculated according to formula and finally drawn：M=27860ng/g dry weights.
Further, recovery testu is carried out using the bulb sample of Lycoris, is carried out under 6 μ g/ml pitch-based spheres
Recovery of standard addition determines, according to step 2-3) method carry out, each level is repeated 3 times, and is as a result measured to containing for lycoramine
Amount is between 20648~27860ng/g (i.e. 0.020~0.027mg/g) dry weight, the rate of recovery 94.8%, RSD 1.0%, table
Bright established extraction and analytical method is accurately and reliably.
Claims (8)
1. a kind of method of lycoramine content in quick detection lycoris plants, including：
(1) lycoris plants sample to be measured is weighed, is pulverized after freeze-drying, using alcohol organic solvent as extractant, is surpassed
Sound extraction, mix, centrifugation, merge supernatant, concentration, then redissolve, be diluted to 1~100ng/ml of concentration, obtain testing sample solution；
(2) testing sample solution is detected using liquid chromatography-tandem second order mses, obtains the peak of lycoramine in sample
Area data；
Wherein, chromatographic condition is as follows：
Liquid-phase chromatographic column：Silica gel type reverse-phase chromatographic column；
Column temperature：40~50 DEG C；
Sample size：5~10 μ L；
Flow velocity：0.2~0.4ml/min；
Mobile phase：Mobile phase A：Formic acid-the aqueous solution, Mobile phase B：Acetonitrile or methanol；
Elution process：The volume ratio of gradient elution, elution time≤6.0min, mobile phase A and Mobile phase B is 40~95：5~
60；
Mass Spectrometry Conditions and scan mode：
Ion gun：Heat electric spray ion source or electric spray ion source；
Ion gun spray voltage：2500~4500V；
Scan mode：With first mass spectrometric full scan-data dependence daughter ion scan pattern or more reaction detections under cation scanning
Pattern is analyzed；
Monitor ion pair：290 ± 1Da of parent ion m/z and at least one lycoramine daughter ion；
Lycoramine retention time：≤3.6min；
(3) according to the standard curve of the peak area data of step (2) and lycoramine obtain sample solution in lycoramine it is dense
Degree, in conjunction with solvent volume and sample dry weight is redissolved in step (1), calculate the content of lycoramine in sample.
2. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：Alcohol organic solvent in the step (1) is methanol or ethanol, and the ratio of sample and extractant is 0.2-1g:2-5ml.
3. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：The ultrasonic extraction time in the step (1) is 15-40min.
4. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：The formic acid content in formic acid-aqueous solution in the step (2) is 0.1~1v/v%.
5. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：The mass-to-charge ratio m/z scopes of daughter ion in the step (2) are 58~290Da.
6. according to claim 1 or 5 in a kind of quick detection lycoris plants lycoramine content method, its feature
It is：The mass-to-charge ratio m/z of the daughter ion is 233 ± 1Da, 215 ± 1Da or 189 ± 1Da.
7. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：Chromatographic condition and Mass Spectrometry Conditions in the step (2) are：40 DEG C of column temperature, sample size 5~10 μ L, 0.2~0.4ml/ of flow velocity
Min, ion gun are heating electric spray ion source, and scan mode is cation Full MS/dd-MS2, ion gun spray voltage
2500V, elution time 6.0min, the retention time of lycoramine is 3.54~3.55min.
8. the method for lycoramine content, its feature exist in a kind of quick detection lycoris plants according to claim 1
In：The preparation method of the standard curve of lycoramine in the step (3) is：Lycoramine standard items are taken, are configured to concentration
1ng/ml~100ng/ml series standard solution, lycoramine standard liquid is entered using liquid chromatography-tandem second order mses
Row measure, standard curve is drawn using the peak area data and lycoramine concentration of standard solution measured, obtains linear equation.
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